Alox12 mediates the temporal shift of arsenite-induced ferroptosis in zebrafish embryonic fibroblast ZF4 cells

Arsenic (As) is a widespread environmental toxicant that poses severe threats to animal and human health by disrupting multiple cellular processes. In this study, embryonic zebrafish fibroblast cells (ZF4) were used as an in vitro model to investigate the role of Alox12-mediated lipoxygenation in arsenite-induced ferroptosis. Transcriptomic analysis revealed that arsenite exposure (20 μM, 24-48 h) globally induced oxidative stress, apoptosis, cell cycle arrest, and autophagic cell death in a time-dependent manner. Notably, ferroptosis was absent at the early stage but became prominent upon prolonged exposure, coinciding with increased Alox12 expression and 12-HETE production. Overexpression of Alox12 under arsenite stress significantly elevated lipid peroxidation, ROS accumulation, and cell death, whereas siRNA-mediated knockdown of Alox12 partially rescued these phenotypes and restored GPx4 activity. These findings demonstrate that Alox12 mediates the transition from ferroptosis suppression to activation during arsenite toxicity, presenting a checkpoint-like phenotypic phenomenon. This work provides mechanistic insight into arsenism and a potential molecular target for ferroptosis-based therapeutic intervention.